#METABOLOMICS WORKBENCH araskind_20170802_132817 DATATRACK_ID:1182 STUDY_ID:ST000833 ANALYSIS_ID:AN001841 PROJECT_ID:PR000594 VERSION 1 CREATED_ON January 16, 2019, 9:11 am #PROJECT PR:PROJECT_TITLE Influence of murine norovirus on cell metabolism PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY C6 effects in RAW PR:INSTITUTE University of Michigan PR:DEPARTMENT Microbiology & Immunology PR:LABORATORY Passalacqua Lab PR:LAST_NAME Passalacqua PR:FIRST_NAME Karla PR:ADDRESS Ann Arbor, MI PR:EMAIL kpassal@umich.edu PR:PHONE 734-763-5639 #STUDY ST:STUDY_TITLE Influence of murine norovirus on cell metabolism ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY 6.5 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to ST:STUDY_SUMMARY recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, ST:STUDY_SUMMARY and 12 mL of medium containing either Murine Norovirus-1 (MNV) at an MOI=5 or ST:STUDY_SUMMARY medium containing a v/v match of mock lysate was added to the cells. Plates were ST:STUDY_SUMMARY rocked on ice for 1 hour, then cells were washed 3X with cold DPBS++, and plain ST:STUDY_SUMMARY medium as added to cells. Plates were then incubated for 7.5 hours at 37 degrees ST:STUDY_SUMMARY and 5% CO2. Cells were washed with 12 mL of 150 mM Ammonium Acetate, swirled 8 ST:STUDY_SUMMARY times, and immediately quenched with liquid nitrogen. Cells were then frozen at ST:STUDY_SUMMARY -80 degrees. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 1 ST:STUDY_COMMENTS BIOHAZARD - MURINE NOROVIRUS - SHOULD BE ENTIRELY NON-INFECTIOUS DUE TO THE ST:STUDY_COMMENTS LIQUID NITROGEN, BUT PLEASE HANDLE AND DISPOSE OF IN BIOHAZARD WASTE. IMPORTANT ST:STUDY_COMMENTS NOTE: I request that the metabolites of the TCA+ assay that are to be assessed ST:STUDY_COMMENTS quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, ST:STUDY_COMMENTS E4P, S7P, 6PG, G3P H6P and Lactate. Also, I would request that both reduced and ST:STUDY_COMMENTS oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the ST:STUDY_COMMENTS non-quantitative data. #SUBJECT SU:SUBJECT_TYPE MOUSE SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0015382 S00023719 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023720 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023721 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023722 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023723 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023724 Medium:MNV infected #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A011_Glycolysis-TCA-nucleotides.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 1mm, 3um) #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE A011_Glycolysis-TCA-nucleotides.pdf AN:ACQUISITION_PARAMETERS_FILE QTOF-002-HILIC-35min-1mm.m #MS MS:INSTRUMENT_NAME Agilent 6520B QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS none #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS uM MS_METABOLITE_DATA_START Samples S00023719 S00023720 S00023721 S00023722 S00023723 S00023724 Factors Medium:mock lysate Medium:mock lysate Medium:mock lysate Medium:MNV infected Medium:MNV infected Medium:MNV infected D-ERYTHROSE 4-PHOSPHATE 2.613316001 9.39460765 6.617425743 2.686075161 3.637145004 2.671098354 BETA-NAD+ .8411 .6508 .9522 .3537 .3948 .8944 COENZYME II .35353663 .270743602 .209430055 .34205688 .254861566 .220849974 SEDOHEPTULOSE 7-PHOSPHATE .294101064 .150612803 .200439074 .340670815 .258101705 .220061428 D-FRUCTOSE, 1,6-BIS(DIHYDROGEN PHOSPHATE) .537716579 .215974879 .240757235 .28408073 .211737232 .215282397 CITRIC ACID 8.839184579 9.457720082 7.454542888 9.680821855 8.786287367 7.032517805 FAD .05256291 .032642277 .018653194 .042580693 .029364375 .029519873 MALIC ACID 5.345311821 1.637526539 .817815478 4.482918214 1.81515342 2.179012735 ADENOSINE TRIPHOSPHATE 70.52576481 23.12110245 100.1014 66.1705 57.4472 74.8022 ROBISON ESTER 2.964311528 1.002351981 1.497850283 2.893330557 1.86805967 1.897242409 SUCROSE 4.553443839 5.817576358 4.736623481 5.671434936 5.745789359 3.81054713 ADENOSINE 5'-DIPHOSPHATE 5.347603573 9.441332401 7.818744872 8.256676598 8.088915322 6.294614524 ADENOSINE 5'-MONOPHOSPHATE 1.378915024 6.550610103 5.286045776 1.709110261 2.03119849 1.695720049 6-PHOSPHOGLUCONIC ACID .217027862 .209660448 .20478464 .21731253 .206793637 .206596261 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type D-ERYTHROSE 4-PHOSPHATE 122357 BETA-NAD+ 15938971 COENZYME II 15938972 SEDOHEPTULOSE 7-PHOSPHATE 165007 D-FRUCTOSE, 1,6-BIS(DIHYDROGEN PHOSPHATE) 21125049 CITRIC ACID 311 FAD 46906035 MALIC ACID 525 ADENOSINE TRIPHOSPHATE 5957 ROBISON ESTER 5958 SUCROSE 5988 ADENOSINE 5'-DIPHOSPHATE 6022 ADENOSINE 5'-MONOPHOSPHATE 6083 6-PHOSPHOGLUCONIC ACID 91493 METABOLITES_END #END